VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression

Autores
Santiago Valtierra, Florencia Ximena; Junco, Milagros; Ventura, Clara; Sanchez Gonzalez, Dativo; Ward, Richard; Garcia Arcos, Itsaso; Hartman, Jessica; Maldonado, Eduardo N.
Año de publicación
2024
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
VDAC1, 2, and 3, are major controllers of mitochondrial metabolism. We previously showed that single and double VDAC knockdown decreased mitochondrial membrane potential (DJm) and that knockdown of the minor isoform VDAC3 decreased cellular ATP and NADH 48 h after transfection in HepG2 hepatocarcinoma cells. Here, we studied the effects of VDAC double knockouts (KO) on mitochondrial metabolism and proliferation. We hypothesized that compensatory mechanisms after VDAC KO induce metabolic changes different from shortterm inhibition of VDAC expression. We used wild-type (wt) and CRISPRCas9 VDAC double KO (VDAC 1-2, VDAC 1-3 and VDAC 2-3 KO) HepG2 cells. We assessed DJm, NADH and mitochondrial morphology by confocal microscopy; respiration by Resipher; triacylglycerides (TG) and cholesterol esters (CE) by thin layer chromatography; and proliferation rate by doubling time DJm, NADH, respiration, mitochondrial morphology, proliferation, and neutral lipids were almost identical in 2-3 VDAC KO cells (VDAC1 present) compared to wt. Absence of VDAC1 (VDAC 1-2/1-3 KO) was associated with changes in metabolism and proliferation. In VDAC 1-2 KO cells (VDAC3 present), DJm and NADH were similar, basal respiration lower, and cell proliferation slower. Both VDAC 1-2 and VDAC 1-3 KO (VDAC2 present) displayed higher TG and CE content than wt. VDAC 1-3 KO cells had similar DJm and respiration, higher NADH and proliferation rates than wt, and giant mitochondria with DJm and NADH 2.0 -fold higher than non-giant counterparts. VDAC1 alone sustains mitochondrial metabolism and proliferation. VDAC1 absence is associated with TG/CE accumulation, and lower rate of proliferation in cells containing only VDAC3, or higher if VDAC2 is the only isoform. These data suggest possible compensatory mechanisms triggered by the lack of specific VDAC isoforms.
Fil: Santiago Valtierra, Florencia Ximena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Medical University of South Carolina; Estados Unidos
Fil: Junco, Milagros. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Medical University of South Carolina; Estados Unidos
Fil: Ventura, Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Grupo Vinculado al IMBICE - Grupo de Biología Estructural y Biotecnología - Universidad Nacional de Quilmes - GBEyB | Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Grupo Vinculado al IMBICE - Grupo de Biología Estructural y Biotecnología - Universidad Nacional de Quilmes - GBEyB | Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular. Grupo Vinculado al IMBICE - Grupo de Biología Estructural y Biotecnología - Universidad Nacional de Quilmes - GBEyB; Argentina. Medical University of South Carolina; Estados Unidos
Fil: Sanchez Gonzalez, Dativo. State University of New York; Estados Unidos
Fil: Ward, Richard. State University of New York; Estados Unidos
Fil: Garcia Arcos, Itsaso. State University of New York; Estados Unidos
Fil: Hartman, Jessica. Medical University of South Carolina; Estados Unidos
Fil: Maldonado, Eduardo N.. Medical University of South Carolina; Estados Unidos
68th Biophysical Society Annual Meeting
San Diego
Estados Unidos
Biophysical Society
Materia
VDAC
MITOCHONDRIA
METABOLISM
CELL PROLIFERATION
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/275716
Acceder
id CONICETDig_d2005ed073cb55079d59fcf4b819bc5d
oai_identifier_str oai:ri.conicet.gov.ar:11336/275716
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expressionSantiago Valtierra, Florencia XimenaJunco, MilagrosVentura, ClaraSanchez Gonzalez, DativoWard, RichardGarcia Arcos, ItsasoHartman, JessicaMaldonado, Eduardo N.VDACMITOCHONDRIAMETABOLISMCELL PROLIFERATIONhttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3VDAC1, 2, and 3, are major controllers of mitochondrial metabolism. We previously showed that single and double VDAC knockdown decreased mitochondrial membrane potential (DJm) and that knockdown of the minor isoform VDAC3 decreased cellular ATP and NADH 48 h after transfection in HepG2 hepatocarcinoma cells. Here, we studied the effects of VDAC double knockouts (KO) on mitochondrial metabolism and proliferation. We hypothesized that compensatory mechanisms after VDAC KO induce metabolic changes different from shortterm inhibition of VDAC expression. We used wild-type (wt) and CRISPRCas9 VDAC double KO (VDAC 1-2, VDAC 1-3 and VDAC 2-3 KO) HepG2 cells. We assessed DJm, NADH and mitochondrial morphology by confocal microscopy; respiration by Resipher; triacylglycerides (TG) and cholesterol esters (CE) by thin layer chromatography; and proliferation rate by doubling time DJm, NADH, respiration, mitochondrial morphology, proliferation, and neutral lipids were almost identical in 2-3 VDAC KO cells (VDAC1 present) compared to wt. Absence of VDAC1 (VDAC 1-2/1-3 KO) was associated with changes in metabolism and proliferation. In VDAC 1-2 KO cells (VDAC3 present), DJm and NADH were similar, basal respiration lower, and cell proliferation slower. Both VDAC 1-2 and VDAC 1-3 KO (VDAC2 present) displayed higher TG and CE content than wt. VDAC 1-3 KO cells had similar DJm and respiration, higher NADH and proliferation rates than wt, and giant mitochondria with DJm and NADH 2.0 -fold higher than non-giant counterparts. VDAC1 alone sustains mitochondrial metabolism and proliferation. VDAC1 absence is associated with TG/CE accumulation, and lower rate of proliferation in cells containing only VDAC3, or higher if VDAC2 is the only isoform. These data suggest possible compensatory mechanisms triggered by the lack of specific VDAC isoforms.Fil: Santiago Valtierra, Florencia Ximena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Medical University of South Carolina; Estados UnidosFil: Junco, Milagros. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Medical University of South Carolina; Estados UnidosFil: Ventura, Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Grupo Vinculado al IMBICE - Grupo de Biología Estructural y Biotecnología - Universidad Nacional de Quilmes - GBEyB | Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Grupo Vinculado al IMBICE - Grupo de Biología Estructural y Biotecnología - Universidad Nacional de Quilmes - GBEyB | Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular. Grupo Vinculado al IMBICE - Grupo de Biología Estructural y Biotecnología - Universidad Nacional de Quilmes - GBEyB; Argentina. Medical University of South Carolina; Estados UnidosFil: Sanchez Gonzalez, Dativo. State University of New York; Estados UnidosFil: Ward, Richard. State University of New York; Estados UnidosFil: Garcia Arcos, Itsaso. State University of New York; Estados UnidosFil: Hartman, Jessica. Medical University of South Carolina; Estados UnidosFil: Maldonado, Eduardo N.. Medical University of South Carolina; Estados Unidos68th Biophysical Society Annual MeetingSan DiegoEstados UnidosBiophysical SocietyCells Press2024info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/275716VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression; 68th Biophysical Society Annual Meeting; San Diego; Estados Unidos; 2024; 525-5251542-0086CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.cell.com/biophysj/abstract/S0006-3495(23)03872-9#articleInformationinfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.bpj.2023.11.3171Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-12-03T09:17:56Zoai:ri.conicet.gov.ar:11336/275716instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-12-03 09:17:57.08CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression
title VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression
spellingShingle VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression
Santiago Valtierra, Florencia Ximena
VDAC
MITOCHONDRIA
METABOLISM
CELL PROLIFERATION
title_short VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression
title_full VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression
title_fullStr VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression
title_full_unstemmed VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression
title_sort VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression
dc.creator.none.fl_str_mv Santiago Valtierra, Florencia Ximena
Junco, Milagros
Ventura, Clara
Sanchez Gonzalez, Dativo
Ward, Richard
Garcia Arcos, Itsaso
Hartman, Jessica
Maldonado, Eduardo N.
author Santiago Valtierra, Florencia Ximena
author_facet Santiago Valtierra, Florencia Ximena
Junco, Milagros
Ventura, Clara
Sanchez Gonzalez, Dativo
Ward, Richard
Garcia Arcos, Itsaso
Hartman, Jessica
Maldonado, Eduardo N.
author_role author
author2 Junco, Milagros
Ventura, Clara
Sanchez Gonzalez, Dativo
Ward, Richard
Garcia Arcos, Itsaso
Hartman, Jessica
Maldonado, Eduardo N.
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv VDAC
MITOCHONDRIA
METABOLISM
CELL PROLIFERATION
topic VDAC
MITOCHONDRIA
METABOLISM
CELL PROLIFERATION
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv VDAC1, 2, and 3, are major controllers of mitochondrial metabolism. We previously showed that single and double VDAC knockdown decreased mitochondrial membrane potential (DJm) and that knockdown of the minor isoform VDAC3 decreased cellular ATP and NADH 48 h after transfection in HepG2 hepatocarcinoma cells. Here, we studied the effects of VDAC double knockouts (KO) on mitochondrial metabolism and proliferation. We hypothesized that compensatory mechanisms after VDAC KO induce metabolic changes different from shortterm inhibition of VDAC expression. We used wild-type (wt) and CRISPRCas9 VDAC double KO (VDAC 1-2, VDAC 1-3 and VDAC 2-3 KO) HepG2 cells. We assessed DJm, NADH and mitochondrial morphology by confocal microscopy; respiration by Resipher; triacylglycerides (TG) and cholesterol esters (CE) by thin layer chromatography; and proliferation rate by doubling time DJm, NADH, respiration, mitochondrial morphology, proliferation, and neutral lipids were almost identical in 2-3 VDAC KO cells (VDAC1 present) compared to wt. Absence of VDAC1 (VDAC 1-2/1-3 KO) was associated with changes in metabolism and proliferation. In VDAC 1-2 KO cells (VDAC3 present), DJm and NADH were similar, basal respiration lower, and cell proliferation slower. Both VDAC 1-2 and VDAC 1-3 KO (VDAC2 present) displayed higher TG and CE content than wt. VDAC 1-3 KO cells had similar DJm and respiration, higher NADH and proliferation rates than wt, and giant mitochondria with DJm and NADH 2.0 -fold higher than non-giant counterparts. VDAC1 alone sustains mitochondrial metabolism and proliferation. VDAC1 absence is associated with TG/CE accumulation, and lower rate of proliferation in cells containing only VDAC3, or higher if VDAC2 is the only isoform. These data suggest possible compensatory mechanisms triggered by the lack of specific VDAC isoforms.
Fil: Santiago Valtierra, Florencia Ximena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Medical University of South Carolina; Estados Unidos
Fil: Junco, Milagros. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Medical University of South Carolina; Estados Unidos
Fil: Ventura, Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Grupo Vinculado al IMBICE - Grupo de Biología Estructural y Biotecnología - Universidad Nacional de Quilmes - GBEyB | Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Grupo Vinculado al IMBICE - Grupo de Biología Estructural y Biotecnología - Universidad Nacional de Quilmes - GBEyB | Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular. Grupo Vinculado al IMBICE - Grupo de Biología Estructural y Biotecnología - Universidad Nacional de Quilmes - GBEyB; Argentina. Medical University of South Carolina; Estados Unidos
Fil: Sanchez Gonzalez, Dativo. State University of New York; Estados Unidos
Fil: Ward, Richard. State University of New York; Estados Unidos
Fil: Garcia Arcos, Itsaso. State University of New York; Estados Unidos
Fil: Hartman, Jessica. Medical University of South Carolina; Estados Unidos
Fil: Maldonado, Eduardo N.. Medical University of South Carolina; Estados Unidos
68th Biophysical Society Annual Meeting
San Diego
Estados Unidos
Biophysical Society
description VDAC1, 2, and 3, are major controllers of mitochondrial metabolism. We previously showed that single and double VDAC knockdown decreased mitochondrial membrane potential (DJm) and that knockdown of the minor isoform VDAC3 decreased cellular ATP and NADH 48 h after transfection in HepG2 hepatocarcinoma cells. Here, we studied the effects of VDAC double knockouts (KO) on mitochondrial metabolism and proliferation. We hypothesized that compensatory mechanisms after VDAC KO induce metabolic changes different from shortterm inhibition of VDAC expression. We used wild-type (wt) and CRISPRCas9 VDAC double KO (VDAC 1-2, VDAC 1-3 and VDAC 2-3 KO) HepG2 cells. We assessed DJm, NADH and mitochondrial morphology by confocal microscopy; respiration by Resipher; triacylglycerides (TG) and cholesterol esters (CE) by thin layer chromatography; and proliferation rate by doubling time DJm, NADH, respiration, mitochondrial morphology, proliferation, and neutral lipids were almost identical in 2-3 VDAC KO cells (VDAC1 present) compared to wt. Absence of VDAC1 (VDAC 1-2/1-3 KO) was associated with changes in metabolism and proliferation. In VDAC 1-2 KO cells (VDAC3 present), DJm and NADH were similar, basal respiration lower, and cell proliferation slower. Both VDAC 1-2 and VDAC 1-3 KO (VDAC2 present) displayed higher TG and CE content than wt. VDAC 1-3 KO cells had similar DJm and respiration, higher NADH and proliferation rates than wt, and giant mitochondria with DJm and NADH 2.0 -fold higher than non-giant counterparts. VDAC1 alone sustains mitochondrial metabolism and proliferation. VDAC1 absence is associated with TG/CE accumulation, and lower rate of proliferation in cells containing only VDAC3, or higher if VDAC2 is the only isoform. These data suggest possible compensatory mechanisms triggered by the lack of specific VDAC isoforms.
publishDate 2024
dc.date.none.fl_str_mv 2024
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/275716
VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression; 68th Biophysical Society Annual Meeting; San Diego; Estados Unidos; 2024; 525-525
1542-0086
CONICET Digital
CONICET
url http://hdl.handle.net/11336/275716
identifier_str_mv VDAC-dependent mitochondrial metabolism and cell proliferation differ between VDAC knockout and transient inhibition of VDAC expression; 68th Biophysical Society Annual Meeting; San Diego; Estados Unidos; 2024; 525-525
1542-0086
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
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info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bpj.2023.11.3171
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
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dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Cells Press
publisher.none.fl_str_mv Cells Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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